Comparative characterization of two monoclonal antibodies targeting canine PD-1

Monoclonal antibodies targeting immune checkpoints have revolutionizedoncology. Yet, the effectiveness of these treatments varies significantly amongpatients, and they are associated with unexpected adverse events, includinghyperprogression. The murine research model used in drug development fails torecapitulate both the functional human immune system and the populationheterogeneity. Hence, a novel model is urgently needed to study theconsequences of immune checkpoint blockade. Dogs appear to be uniquelysuited for this role. Approximately 1 in 4 companion dogs dies from cancer, yetno antibodies are commercially available for use in veterinary oncology. Here wecharacterize two novel antibodies that bind canine PD-1 with sub-nanomolaraffinity as measured by SPR. Both antibodies block the clinically crucial PD-1/PDL1 interaction in a competitive ELISA assay. Additionally, the antibodies weretested with a broad range of assays including Western Blot, ELISA, flowcytometry, immunofluorescence and immunohistochemistry. The antibodiesappear to bind two distinct epitopes as predicted by molecular modeling andpeptide phage display. Our study provides new tools for canine oncologyresearch and a potential veterinary therapeutic

Biochemical evidence for conformational variants in the anti-viral and pro-metastatic protein IFITM1

Interferon induced transmembrane proteins (IFITMs) play a dual role in the restriction of RNA viruses and in cancer progression, yet the mechanism of their action remains unknown. Currently, there is no data about the basic biochemical features or biophysical properties of the IFITM1 protein. In this work, we report on description and biochemical characterization of three conformational variants/oligomeric species of recombinant IFITM1 protein derived from an E. coli expression system. The protein was extracted from the membrane fraction, affinity purified, and separated by size exclusion chromatography where two distinct oligomeric species were observed in addition to the expected monomer. These species remained stable upon re-chromatography and were designated as “dimer” and “oligomer” according to their estimated molecular weight. The dimer was found to be less stable compared to the oligomer using circular dichroism thermal denaturation and incubation with a reducing agent. A two-site ELISA and HDX mass spectrometry suggested the existence of structural motif within the N-terminal part of IFITM1 which might be significant in oligomer formation. Together, these data show the unusual propensity of recombinant IFITM1 to naturally assemble into very stable oligomeric species whose study might shed light on IFITM1 anti-viral and pro-oncogenic functions in cells

Two-level Control of Biological Activation in Waste-treatment Plant

Tato diplomová práce se zabývá návrhem a řízením dvoustupňové regulace biologické aktivace čistírny odpadních vod v Havířově. Praktická část a přínos této práce spočívá především v přenesení výsledků přímo do praxe, kde byla jednostupňová regulace biologické aktivace nahrazena dvoustupňovou a sestava TC 700 byla rozšířena o modul DataBoxu pro vyčítání a zpracování ukládaných dat.This dissertation deals with the proposal and operating of the two-level Control of Biological Activation in Waste-treatment Plant in Havířov. The practical part and a contribution of this work consists mainly in the translation of the results directly to the practice, where the one-level Control of Biological Activation has been replaced by the two-level Control of Biological Activation and the configuration TC 700 was extended by the modulus DataBox for twitting and processing of the storing data.Prezenční455 - Katedra měřicí a řídicí technikyvýborn